Attention:

Certain features of Sigma-Aldrich.com will be down for maintenance the evening of Friday August 18th starting at 8:00 pm CDT until Saturday August 19th at 12:01 pm CDT. Please note that you still have telephone and email access to our local offices. We apologize for any inconvenience.

EnPresso® B Animal-free

Description

EnPresso B is a pre-sterilized growth system designed to increase the yield of functional protein from E. coli-based expression systems. EnPresso growth systems provide optimal conditions for growth, metabolism and protein expression in microbial cultures. Protein yields are increased by enabling cultures to reach far higher cell densities than those achieved using conventional media. By controlling growth rate and metabolism, a greater proportion of expressed protein can be correctly folded to improve solubility, minimize the risk of inclusion body formation, and ensure functionality of the final product. EnPresso growth systems maintain pH, provide adequate minerals, vitamins and trace elements to support growth, and use proprietary EnBase® technology to ensure a constant, slow release of glucose from a polysaccharide substrate.

Additional items needed

Anti-foaming agent, such as AntiFoam 204, when using Ultra Yield Flasks or other baffled (fluted) flasks

Note: To improve aeration in shake flask cultures, we recommend the use of Ultra Yield Shake Flasks with AirOtop Enhanced Seals. Improved aeration has been shown to further enhance the performance of EnPresso growth systems for bacterial cultures.

Recommended conditions

Culture volume: 10% of flask volume (20% only if using Ultra Yield flasks)

IMPORTANT NOTICE:
It is essential to follow EnPresso B Animal-free protocols in detail. Using a fresh pre-culture, the correct shake speed, air-permeable closures and recommended cultivation times are critical to ensure success. Never use aluminum foil or plastic lids during incubation.

Protocol for shake flasks – 50 mL culture

Day 1 - step 1

Prepare inoculum from a glycerol stock or use a single colony grown overnight on an agar plate. Inoculate 2 mL of LB medium containing antibiotics in a cultivation tube or shake flask.

Incubate at 37 °C for 6-8 hours with vigorous shaking.

Day 1 - step 2

Add contents of one white bag (2 tablets) to 50 mL of sterile water in a sterile 500 mL shake flask.